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A dual signal amplification strategy combining thermally initiated SI-RAFT polymerization and DNA-templated silver nanoparticles for electrochemical determination of DNA

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Abstract

A highly sensitive method is described for determination of DNA. It is based on dual signal amplification, viz. (a)DNA-templated metal deposition, and (b) thermally initiated surface-initiated reversible addition−fragmentation chain transfer (SI-RAFT) polymerization. A peptide nucleic acid (PNA) with a terminal thiol group was grasped onto a gold electrode by self-assembly. The modified electrode serves as a probe to selectively capture target DNA (tDNA). In the next step, Zr(IV) ions are bound to the phosphate groups of the tDNA. A chain-transfer agent (CTA) for thermally initiated SI-RAFT polymerization, 4-cyano-4-(phenylcarbonothioylthio)pentanoic acid (CPAD), was immobilized on tDNA by conjugation of the carboxy group to Zr(IV) ions. Subsequently, numerous monomers of glycosyloxyethyl methacrylate (GEMA) were connected to the CPAD by thermally initiated SI-RAFT polymerization with azobisisobutyronitrile (AIBN) serving as the free-radical thermal initiator. Afterwards, hydroxyl groups of the GEMA were oxidized to aldehyde groups reacting with sodium periodate, and silver nanoparticles were further introduced on the surface of electrode via “silver mirror reaction”. This results in a large electrochemical signal amplification. Under optimized conditions, the electrochemical signal (best measured at a working potential of 0 V vs. SCE (KCl; 3 M)) increases linearly with the logarithm of tDNA concentration in the 10 to 106 aM concentration range. The detection limit is as low as 5.6 aM (~34 molecules in a 10 μL sample). This is lower by factors between 2 and 1800 times than detection limits of most other ultra-sensitive electrochemical DNA assays.

Schematic representation of a dual signal amplification strategy combining thermally initiated surface-initiated reversible addition−fragmentation chain transfer polymerization (SI-RAFT) and DNA-templated silver nanoparticles for electrochemical determination of DNA.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 21575066 and 21974068).

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Authors and Affiliations

  1. School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, Jiangsu, 210094, People’s Republic of China

    Bang Liu, Haobo Sun & Jinming Kong

  2. Laboratory of Translational Medicine, Jiangsu Province Academy of Traditional Chinese Medicine, No.100, Shizi Street, Hongshan Road, Qixia District, Nanjing, 210028, Jiangsu Province, People’s Republic of China

    Bang Liu & Jian Zhang

  3. School of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, 252059, China

    Lianzhi Li

  4. School of Biomedical Engineering, Shenzhen University Health Science Center, Shenzhen, Guangdong, 518060, People’s Republic of China

    Xueji Zhang

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  1. Bang Liu
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  4. Jian Zhang
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Correspondence to Jian Zhang or Jinming Kong.

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Liu, B., Sun, H., Li, L. et al. A dual signal amplification strategy combining thermally initiated SI-RAFT polymerization and DNA-templated silver nanoparticles for electrochemical determination of DNA. Microchim Acta 187, 35 (2020). https://doi.org/10.1007/s00604-019-3912-9

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